title: Genes required for and effects of alginate overproduction induced by growth of Pseudomonas aeruginosa on Pseudomonas isolation agar supplemented with ammonium metavanadate.
abstract: Pseudomonas aeruginosa is an opportunistic pathogen that can adapt to changing environments and can secrete an exopolysaccharide known as alginate as a protection response, resulting in a colony morphology and phenotype referred to as mucoid. However, how P. aeruginosa senses its environment and activates alginate overproduction is not fully understood. Previously, we showed that Pseudomonas isolation agar supplemented with ammonium metavanadate (PIAAMV) induces P. aeruginosa to overproduce alginate. Vanadate is a phosphate mimic and causes protein misfolding by disruption of disulfide bonds. Here we used PIAAMV to characterize the pathways involved in inducible alginate production and tested the global effects of P. aeruginosa growth on PIAAMV by a mutant library screen, by transcriptomics, and in a murine acute virulence model. The PA14 nonredundant mutant library was screened on PIAAMV to identify new genes that are required for the inducible alginate stress response. A functionally diverse set of genes encoding products involved in cell envelope biogenesis, peptidoglycan remodeling, uptake of phosphate and iron, phenazine biosynthesis, and other processes were identified as positive regulators of the mucoid phenotype on PIAAMV. Transcriptome analysis of P. aeruginosa cultures growing in the presence of vanadate showed differential expression of genes involved in virulence, envelope biogenesis, and cell stress pathways. In this study, it was observed that growth on PIAAMV attenuates P. aeruginosa in a mouse pneumonia model. Induction of alginate overproduction occurs as a stress response to protect P. aeruginosa, but it may be possible to modulate and inhibit these pathways based on the new genes identified in this study.
authors.affiliation: Department of Microbiology, Immunology, and Cancer Biology, University of Virginia Health System, Charlottesville, VA, USA.
authors.forename: F Heath
authors.initials: FH
authors.lastname: Damron
authors.forename: Mariette
authors.initials: M
authors.lastname: Barbier
authors.forename: Elizabeth S
authors.initials: ES
authors.lastname: McKenney
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Schurr
authors.forename: Joanna B
authors.initials: JB
authors.lastname: Goldberg
mesh_terms: D000208:Acute Disease; D000362:Agar; D000464:Alginates / Q000378:metabolism*; D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D003470:Culture Media / Q000737:chemistry; D004195:Disease Models, Animal; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D020723:Glucuronic Acid / Q000378:metabolism; D018869:Heat-Shock Response; D006603:Hexuronic Acids / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D018410:Pneumonia, Bacterial / Q000382:microbiology; D011552:Pseudomonas Infections / Q000382:microbiology; D011550:Pseudomonas aeruginosa / Q000187:drug effects* / Q000235:genetics / Q000254:growth & development* / Q000472:pathogenicity; D014638:Vanadates / Q000378:metabolism / Q000494:pharmacology*; D014774:Virulence / Q000235:genetics
chemical_list: D000464:Alginates; D001426:Bacterial Proteins; D003470:Culture Media; D006603:Hexuronic Acids; D014638:Vanadates; D020723:Glucuronic Acid; D000362:Agar; C101036:ammonium metavanadate